Neuroinflammation and oxidative stress are deemed the prime causes of brain injury after cerebral ischemia/reperfusion (I/R). Since the silent mating-type information regulation 2 homologue 3 (Sirt3) pathway plays an imperative role in protecting against neuroinflammation and oxidative stress, it has been verified as a target to treat ischemia stroke. Therefore, we attempted to seek novel Sirt3 agonist and explore its underlying mechanism for stroke treatment both in vivo and in vitro.
Results:
Trilobatin (TLB) not only dramatically suppressed neuroinflammation and oxidative stress injury after middle cerebral artery occlusion in rats, but also effectively mitigated oxygen and glucose deprivation/reoxygenation injury in primary cultured astrocytes. These beneficial effects, along with the reduced proinflammatory cytokines via suppressing Toll-like receptor 4 (TLR4) signaling pathway, lessened oxidative injury via activating nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathways, in keeping with the findings in vivo. Intriguingly, the TLB-mediated neuroprotection on cerebral I/R injury was modulated by reciprocity between TLR4-mediated neuroinflammatory responses and Nrf2 antioxidant responses as evidenced by molecular docking and silencing TLR4 and Nrf2, respectively. Most importantly, TLB not only directly bonded to Sirt3 but also increased Sirt3 expression and activity, indicating that Sirt3 might be a promising therapeutic target of TLB.
Innovation:
TLB is a naturally occurring Sirt3 agonist with potent neuroprotective effects via regulation of TLR4/nuclear factor-kappa B and Nrf2/Kelch-like ECH-associated protein 1 (Keap-1) signaling pathways both in vivo and in vitro.
Conclusion:
Our findings indicate that TLB protects against cerebral I/R-induced neuroinflammation and oxidative injury through the regulation of neuroinflammatory and oxidative responses via TLR4, Nrf2, and Sirt3, suggesting that TLB might be a promising Sirt3 agonist against ischemic stroke.
Get full access to this article
View all access options for this article.
References
1.
AlfieriA, SrivastavaS, SiowRCM, CashD, ModoM, DuchenMR, FraserPA, WilliamsSCR, and MannGE. Sulforaphane preconditioning of the Nrf2/HO-1 defense pathway protects the cerebral vasculature against blood-brain barrier disruption and neurological deficits in stroke. Free Radic Biol Med, 65: 1012–1022, 2013.
2.
AlimI, CaulfieldJT, ChenY, SwarupV, GeschwindDH, IvanovaE, SeravalliJ, AiY, SansingLH, Ste MarieEJ, HondalRJ, MukherjeeS, CaveJW, SagdullaevBT, KaruppagounderSS, and RatanRR. Selenium drives a transcriptional adaptive program to block ferroptosis and treat stroke. Cell, 177: 1262–1279.e25, 2019.
3.
BachstetterAD, WebsterSJ, GouldingDS, MortonJE, WattersonDM, and Van EldikLJ. Attenuation of traumatic brain injury-induced cognitive impairment in mice by targeting increased cytokine levels with a small molecule experimental therapeutic. J Neuroinflammation, 12: 69, 2015.
4.
ChenT, DaiSH, LiX, LuoP, ZhuJ, WangYH, FeiZ, and JiangXF. Sirt1-Sirt3 axis regulates human blood-brain barrier permeability in response to ischemia. Redox Biol, 14: 229–236, 2018.
5.
Collaborators GBDCoD. Global, regional, and national age-sex-specific mortality for 282 causes of death in 195 countries and territories, 1980–2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet, 392: 1736–1788, 2018.
6.
FanX, ZhangY, DongH, WangB, JiH, and LiuX. Trilobatin attenuates the LPS-mediated inflammatory response by suppressing the NF-kappaB signaling pathway. Food Chem, 166: 609–615, 2015.
7.
GaoJ, DengY, YinC, LiuY, ZhangW, ShiJ, and GongQ. Icariside II, a novel phosphodiesterase 5 inhibitor, protects against H2O2-induced PC12 cells death by inhibiting mitochondria-mediated autophagy. J Cell Mol Med, 21: 375–386, 2017.
8.
GaoJ, LiuS, XuF, LiuY, LvC, DengY, ShiJ, and GongQ. Trilobatin protects against oxidative injury in neuronal PC12 cells through regulating mitochondrial ROS homeostasis mediated by AMPK/Nrf2/Sirt3 signaling pathway. Front Mol Neurosci, 11: 267, 2018.
9.
GuptaS and GuptaYK. Combination of Zizyphus jujuba and silymarin showed better neuroprotective effect as compared to single agent in MCAo-induced focal cerebral ischemia in rats. J Ethnopharmacol, 197: 118–127, 2017.
10.
HindWH, TufarelliC, NeophytouM, AndersonSI, EnglandTJ, and O'SullivanSE. Endocannabinoids modulate human blood-brain barrier permeability in vitro. Br J Pharmacol, 172: 3015–3027, 2015.
11.
IkramM, MuhammadT, RehmanSU, KhanA, JoMG, AliT, and KimMO. Hesperetin confers neuroprotection by regulating Nrf2/TLR4/NF-kappaB signaling in an Abeta mouse model. Mol Neurobiol, 56: 6293–6309, 2019.
12.
KahlesT and BrandesRP. Which NADPH oxidase isoform is relevant for ischemic stroke? The case for nox 2. Antioxid Redox Signal, 18: 1400–1417, 2013.
13.
KuangX, WangLF, YuL, LiYJ, WangYN, HeQ, ChenC, and DuJR. Ligustilide ameliorates neuroinflammation and brain injury in focal cerebral ischemia/reperfusion rats: involvement of inhibition of TLR4/peroxiredoxin 6 signaling. Free Radic Biol Med, 71: 165–175, 2014.
14.
LiuL, VollmerMK, AhmadAS, FernandezVM, KimH, and DoreS. Pretreatment with Korean red ginseng or dimethyl fumarate attenuates reactive gliosis and confers sustained neuroprotection against cerebral hypoxic-ischemic damage by an Nrf2-dependent mechanism. Free Radic Biol Med, 131: 98–114, 2019.
15.
LiuT, MaX, OuyangT, ChenH, XiaoY, HuangY, LiuJ, and XuM. Efficacy of 5-aminolevulinic acid-based photodynamic therapy against keloid compromised by downregulation of SIRT1-SIRT3-SOD2-mROS dependent autophagy pathway. Redox Biol, 20: 195–203, 2019.
16.
LongaEZ, WeinsteinPR, CarlsonS, and CumminsR. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke, 20: 84–91, 1989.
17.
LudewigP, WinnebergerJ, and MagnusT. The cerebral endothelial cell as a key regulator of inflammatory processes in sterile inflammation. J Neuroimmunol, 326: 38–44, 2019.
18.
LuoY, WangC, LiWH, LiuJ, HeHH, LongJH, YangJ, SuiX, WangS, YouZ, and WangYA. Madecassoside protects BV2 microglial cells from oxygen-glucose deprivation/reperfusion-induced injury via inhibition of the toll-like receptor 4 signaling pathway. Brain Res, 1679: 144–154, 2018.
NalamoluKR, SmithNJ, ChelluboinaB, KlopfensteinJD, PinsonDM, WangDZ, VemugantiR, and VeeravalliKK. Prevention of the severity of post-ischemic inflammation and brain damage by simultaneous knockdown of toll-like receptors 2 and 4. Neuroscience, 373: 82–91, 2018.
21.
NarayananSV, DaveKR, SaulI, and Perez-PinzonMA. Resveratrol preconditioning protects against cerebral ischemic injury via nuclear erythroid 2-related factor 2. Stroke, 46: 1626–1632, 2015.
22.
NashKM, SchieferIT, and ShahZA. Development of a reactive oxygen species-sensitive nitric oxide synthase inhibitor for the treatment of ischemic stroke. Free Radic Biol Med, 115: 395–404, 2018.
23.
ParadaE, CasasAI, Palomino-AntolinA, Gomez-RangelV, Rubio-NavarroA, Farre-AlinsV, Narros-FernandezP, Guerrero-HueM, MorenoJA, RosaJM, RodaJM, Hernandez-GarciaBJ, and EgeaJ. Early TLR4 blockade reduces ROS and inflammation triggered by microglial pro-inflammatory phenotype in rodents and human brain ischemia models. Br J Pharmacol, 176: 2764–2779, 2019.
24.
PengL, YinJ, GeM, WangS, XieL, LiY, SiJQ, and MaK. Isoflurane post-conditioning ameliorates cerebral ischemia/reperfusion injury by enhancing angiogenesis through activating the Shh/Gli signaling pathway in rats. Front Neurosci, 13: 321, 2019.
25.
PinchukI, WeberD, KochlikB, StuetzW, ToussaintO, Debacq-ChainiauxF, DolleMET, JansenE, GonosES, SikoraE, BreusingN, GradinaruD, SindlingerT, Moreno-VillanuevaM, BurkleA, GruneT, and LichtenbergD. Gender- and age-dependencies of oxidative stress, as detected based on the steady state concentrations of different biomarkers in the MARK-AGE study. Redox Biol, 24: 101204, 2019.
26.
PuH, ShiY, ZhangL, LuZ, YeQ, LeakRK, XuF, MaS, MuH, WeiZ, XuN, XiaY, HuX, HitchensTK, BennettMVL, and ChenJ. Protease-independent action of tissue plasminogen activator in brain plasticity and neurological recovery after ischemic stroke. Proc Natl Acad Sci U S A, 116: 9115–9124, 2019.
27.
SivandzadeF, PrasadS, BhaleraoA, and CuculloL. NRF2 and NF-B interplay in cerebrovascular and neurodegenerative disorders: molecular mechanisms and possible therapeutic approaches. Redox Biol, 21: 101059, 2019.
28.
SongS, WangS, PigottVM, JiangT, FoleyLM, MishraA, NayakR, ZhuW, BegumG, ShiY, CarneyKE, HitchensTK, ShullGE, and SunD. Selective role of Na+/H+ exchanger in Cx3cr1+ microglial activation, white matter demyelination, and post-stroke function recovery. Glia, 66: 2279–2298, 2018.
29.
TrabaJ, GeigerSS, Kwarteng-SiawM, HanK, RaOH, SiegelRM, GiusD, and SackMN. Prolonged fasting suppresses mitochondrial NLRP3 inflammasome assembly and activation via SIRT3-mediated activation of superoxide dismutase 2. J Biol Chem, 292: 12153–12164, 2017.
30.
VartanianKB, StevensSL, MarshBJ, Williams-KarneskyR, LessovNS, and Stenzel-PooreMP. LPS preconditioning redirects TLR signaling following stroke: TRIF-IRF3 plays a seminal role in mediating tolerance to ischemic injury. J Neuroinflammation, 8: 140, 2011.
31.
WangJ, ShiY, ZhangL, ZhangF, HuX, ZhangW, LeakRK, GaoY, ChenL, and ChenJ. Omega-3 polyunsaturated fatty acids enhance cerebral angiogenesis and provide long-term protection after stroke. Neurobiol Dis, 68: 91–103, 2014.
32.
WangQ, GongQ, WuQ, and ShiJ. Neuroprotective effects of Dendrobium alkaloids on rat cortical neurons injured by oxygen-glucose deprivation and reperfusion. Phytomedicine, 17: 108–115, 2010.
33.
WangY, HuangY, XuY, RuanW, WangH, ZhangY, SaavedraJM, ZhangL, HuangZ, and PangT. A dual AMPK/Nrf2 activator reduces brain inflammation after stroke by enhancing microglia M2 polarization. Antioxid Redox Signal, 28: 141–163, 2018.
34.
WangY, JinH, WangW, WangF, and ZhaoH. Myosin1f-mediated neutrophil migration contributes to acute neuroinflammation and brain injury after stroke in mice. J Neuroinflammation, 16: 77, 2019.
35.
YagiK, KitazatoKT, UnoM, TadaY, KinouchiT, ShimadaK, and NagahiroS. Edaravone, a free radical scavenger, inhibits MMP-9-related brain hemorrhage in rats treated with tissue plasminogen activator. Stroke, 40: 626–631, 2009.
36.
YangL, ZhangXY, LiK, LiAP, YangWD, YangR, WangP, ZhaoZH, CuiF, QinY, YangJH, TaoHL, SunT, ChenS, YuPH, LiuHJ, and YangC. Protopanaxadiol inhibits epithelial-mesenchymal transition of hepatocellular carcinoma by targeting STAT3 pathway. Cell Death Dis, 10: 630, 2019.
37.
YangT, SunY, MaoL, ZhangM, LiQ, ZhangL, ShiY, LeakRK, ChenJ, and ZhangF. Brain ischemic preconditioning protects against ischemic injury and preserves the blood-brain barrier via oxidative signaling and Nrf2 activation. Redox Biol, 17: 323–337, 2018.
ZengJ, ChenY, DingR, FengL, FuZ, YangS, DengX, XieZ, and ZhengS. Isoliquiritigenin alleviates early brain injury after experimental intracerebral hemorrhage via suppressing ROS- and/or NF-κB-mediated NLRP3 inflammasome activation by promoting Nrf2 antioxidant pathway. J Neuroinflammation, 14: 119, 2017.
42.
ZhangG, ZhangT, LiN, WuL, GuJ, LiC, ZhaoC, LiuW, ShanL, YuP, YangX, TangY, YangGY, WangY, SunY, and ZhangZ. Tetramethylpyrazine nitrone activates the BDNF/Akt/CREB pathway to promote post-ischaemic neuroregeneration and recovery of neurological functions in rats. Br J Pharmacol, 175: 517–531, 2018.
43.
ZhangK, ZhangQ, DengJ, LiJ, LiJ, WenL, MaJ, and LiC. ALK5 signaling pathway mediates neurogenesis and functional recovery after cerebral ischemia/reperfusion in rats via Gadd45b. Cell Death Dis, 10: 360, 2019.
44.
ZhangL, SeoJH, LiH, NamG, and YangHO. The phosphodiesterase 5 inhibitor, KJH-1002, reverses a mouse model of amnesia by activating a cGMP/cAMP response element binding protein pathway and decreasing oxidative damage. Br J Pharmacol, 175: 3347–3360, 2018.
45.
ZhangM, MaY, ChaiL, MaoH, ZhangJ, and FanX. Storax protected oxygen-glucose deprivation/reoxygenation induced primary astrocyte injury by inhibiting NF-κB activation in vitro. Front Pharmacol, 9: 1527, 2018.
46.
ZhangQ, WuHH, WangY, GuGJ, ZhangW, and XiaR. Neural stem cell transplantation decreases neuroinflammation in a transgenic mouse model of Alzheimer's disease. J Neurochem, 136: 815–825, 2016.
47.
ZhangW, LiuJ, HuX, LiP, LeakRK, GaoY, and ChenJ. n-3 Polyunsaturated fatty acids reduce neonatal hypoxic/ischemic brain injury by promoting phosphatidylserine formation and Akt signaling. Stroke, 46: 2943–2950, 2015.
48.
ZhangX, TangX, LiuK, HamblinMH, and YinKJ. Long Noncoding RNA Malat1 regulates cerebrovascular pathologies in ischemic stroke. J Neurosci, 37: 1797–1806, 2017.
49.
ZhangZ, QinP, DengY, MaZ, GuoH, GuoH, HouY, WangS, ZouW, SunY, MaY, and HouW. The novel estrogenic receptor GPR30 alleviates ischemic injury by inhibiting TLR4-mediated microglial inflammation. J Neuroinflammation, 15: 206, 2018.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
